Development of a cofactor balanced, multi enzymatic cascade reaction for the simultaneous production of l-alanine and l-serine from 2-keto-3-deoxy-gluconate

Benjamin Begander, Anna Huber, Josef Sperl, Volker Sieber

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Enzymatic reaction cascades represent a powerful tool to convert biogenic resources into valuable chemicals for fuel and commodity markets. Sugars and their breakdown products constitute a significant group of possible substrates for such biocatalytic conversion strategies to value-added products. However, one major drawback of sugar cascades is the need for cofactor recycling without using additional enzymes and/or creating unwanted by-products. Here, we describe a novel, multi-enzymatic reaction cascade for the one-pot simultaneous synthesis of L-alanine and L-serine, using the sugar degradation product 2-keto-3-deoxygluconate and ammonium as precursors. To pursue this aim, we used four different, thermostable enzymes, while the necessary cofactor NADH is recycled entirely self-sufficiently. Buffer and pH optimisation in combination with an enzyme titration study yielded an optimised production of 21.3 +/− 1.0 mM L-alanine and 8.9 +/− 0.4 mM L-serine in one pot after 21 h.

Original languageEnglish
Article number31
Pages (from-to)1-12
Number of pages12
JournalCatalysts
Volume11
Issue number1
DOIs
StatePublished - Jan 2021

Keywords

  • Amino acids
  • Cofactor self-sufficient
  • Enzyme reaction
  • Multi-enzyme cascade reaction
  • One-pot synthesis
  • Optimisation

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